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Targeting agent for cancer cell or cancer-associated fibroblast

a cancer cell or cancer-associated fibroblast technology, applied in the direction of viruses, drug compositions, dna/rna fragmentation, etc., can solve the problems of nausea and vomiting, hair loss, myelosuppression, and no delivery method is satisfactory, etc., to suppress the activity or growth, suppress the advance, and efficiently deliver to

Active Publication Date: 2014-04-01
NITTO DENKO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0031]The carrier of the present invention specifically targets a cancer cell and a CAF, and efficiently delivers to a cancer cell and / or a CAF a desired substance or body such as, for example, a drug that controls the activity or growth of a cancer cell or a CAF, thus enabling a desired effect such as, for example, suppression of the activity or growth of a cancer cell or a CAF thereby curing cancer, suppressing the advance thereof, and preventing the onset thereof, to be achieved with the highest efficiency and the minimum side effects.
[0032]Since the anticancer composition of the present invention is based on the completely novel approach of treating a cancer by acting on a CAF in addition to a cancer cell itself, efficacy can be expected on cancers for which a conventional treatment method could not give satisfactory results and, furthermore, a synergistic effect due to combined use with a conventional anticancer agent, angiogenesis inhibitor, etc. can be anticipated.
[0033]Furthermore, since the carrier of the present invention can specifically deliver a substance to a cancer cell and a CAF, it can be utilized for specifically labeling a cancer cell and a CAF, gene transfer, etc.

Problems solved by technology

In cancer treatment, particularly in the medical therapy of cancer, various anticancer agents for suppressing the growth of cancer cells have been developed, and some degree of success has been achieved, but since such drugs suppress the growth of not only cancer cells but also normal cells, there are problems with various side effects such as nausea and vomiting, hair loss, myelosuppression, kidney damage, and nerve damage.
However, none of the delivery methods are yet satisfactory, and there has been a further desire for the development of cancer cell-specific delivery methods.
However, such attempts could not always achieve satisfactory results because of the above-mentioned problems with side effects, or the occurrence of additional phenomena such as relapse due to minimal residual disease, resistance of tumor cells to the anticancer agent, etc.

Method used

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  • Targeting agent for cancer cell or cancer-associated fibroblast
  • Targeting agent for cancer cell or cancer-associated fibroblast
  • Targeting agent for cancer cell or cancer-associated fibroblast

Examples

Experimental program
Comparison scheme
Effect test

example 1

Separation of CAFs

[0109]Cancer tissue or peripheral normal tissue (normal tissue separated from a site spaced from cancer tissue by at least 2 cm) removed from a colon cancer patient was finely cut into 1×1×1 mm, then centrifugally washed with PBS twice, and the pellets were cultured in a culture liquid (DMEM (Dulbecco's Modified Eagle Medium) containing collagenase type I (225 U / ml), hyaluronidase (125 U / ml), 10% FBS (fetal bovine serum), streptomycin / penicillin) for 24 hours. Subsequently, the supernatant was aspirated, and culturing was continued after changing the liquid culture for 10% FBS / DMEM. When the cultured cells were immunostained with an FITC labeled antibody with respect to α-SMA, which is a marker for CAFs, and vimentin, which is a marker for mesenchymal cells, α-SMA was detected only in cancer tissue-derived cells, and it was confirmed that these cells were CAFs (see FIG. 1). Vimentin was positive for cells derived from either tissue, and desmin, which is a marker fo...

example 2

CAF Tumor Growth Activity

[0110]A 6-well plate was seeded with CAFs or normal fibroblasts obtained in Example 1 at a density of 1×105 cells / well and cultured with 10% FBS / DMEM, the liquid culture was replaced with 0.5% FBS / DMEM in a confluent state on the third day, and the liquid culture was seeded with colon cancer cell line M7609 cells (2×105 cells), and coculturing was carried out for 7 days. The number of M7609 cells was counted with a Coulter counter (Beckman) at 0 days and on the 3rd and 5th days. The results are given in FIG. 2. This shows that CAFs promote the growth of cancer cells.

example 3

Preparation of siRNA

[0111]Three types of siRNA targeted at gp46 (GenBank Accession No. M69246), which is a rat homologue of human HSP47, and a random siRNA control were purchased from Hokkaido System Science Co., Ltd. Each siRNA consists of 27 bases overhanging on the 3′ side, and the sequences are as follows.

[0112]

Sequence A:(sense, SEQ ID NO: 1)5′-GUUCCACCAUAAGAUGGUAGACAACAG-3′(antisense, SEQ ID NO: 2)5′-GUUGUCUACCAUCUUAUGGUGGAACAU-3′Sequence B:(sense, SEQ ID NO: 3)5′-CCACAAGUUUUAUAUCCAAUCUAGCAG-3′(antisense, SEQ ID NO: 4)5′-GCUAGAUUGGAUAUAAAACUUGUGGAU-3′Sequence C:(sense, SEQ ID NO: 5)5′-CUAGAGCCAUUACAUUACAUUGACAAG-3′(antisense, SEQ ID NO: 6)5′-UGUCAAUGUAAUGUAAUGGCUCUAGAU-3′Random siRNA:(sense, SEQ ID NO: 7)5′-CGAUUCGCUAGACCGGCUUCAUUGCAG-3′(antisense, SEQ ID NO: 8)5′-GCAAUGAAGCCGGUCUAGCGAAUCGAU-3′

[0113]Furthermore, siRNA labeled on the 5′ side with the fluorescent dye 6′-carboxyfluorescein (6-FAM) was also prepared.

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Abstract

Disclosed are a novel therapeutic agent and a novel treatment method for cancer. Specifically disclosed are: a targeting agent for a cell selected from the group consisting of a cancer cell and a cancer-associated fibroblast, which comprises a retinoid; a substance delivery carrier for the cell, which comprises the targeting agent; an anti-cancer composition utilizing the targeting agent or the carrier; an anti cancer-associated fibroblast composition; and a method for treatment of cancer.

Description

TECHNICAL FIELD[0001]The present invention relates to a targeting agent to a cell selected from the group consisting of a cancer cell and a cancer-associated fibroblast (CAF: cancer-associated fibroblast or carcinoma-associated fibroblast), a substance delivery carrier to the cell, the carrier containing the targeting agent, and an anticancer composition, an anti-CAF composition, and a method for treating a cancer utilizing same.BACKGROUND ART[0002]Cancer is one of the most significant diseases confronting mankind, and much research effort is going into the treatment thereof. In cancer treatment, particularly in the medical therapy of cancer, various anticancer agents for suppressing the growth of cancer cells have been developed, and some degree of success has been achieved, but since such drugs suppress the growth of not only cancer cells but also normal cells, there are problems with various side effects such as nausea and vomiting, hair loss, myelosuppression, kidney damage, and...

Claims

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Application Information

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Patent Type & Authority Patents(United States)
IPC IPC(8): A61K48/00A61K31/07A61K9/127C07C35/18A01N31/04C12N15/113C12N15/11
CPCA61K31/704A61K48/005C12N15/88C12N2810/10A61K48/00A61K9/127A61K2300/00C12N2320/32A61K31/713C12N15/113C12N2310/14C12N2799/04A61K9/0019A61K31/07A61K47/48815A61K47/10A61K47/6911A61P35/00A61P35/02A61P43/00A61K31/203
Inventor NIITSU, YOSHIROTAKIMOTO, RISHU
Owner NITTO DENKO CORP
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